Fig. 10

Ru1 inhibits mtDNA transcription, OXPHOS protein complex translation and mitochondrial mass. A Mean fold-change ± SD in the expression of the mtDNA-encoded gene MTATP6, MTCYTB or MTND4 and the nuclear DNA encoded gene COX5 in Panc354 tumors extracted on d19 from mice treated with Ctl or Ru1 (1.4mg/kg; daily, r.o.). Values were normalized to ß-actin levels. * p < 0.05, ** p < 0.01, *** p<0.001, ns = not significant, as determined by unpaired two-sided Student’s t-test. B Mean fold-change ± SEM in the expression of the mtDNA-encoded gene MTATP6 or the nuclear DNA encoded gene COX5 in Panc265 orthotopic tumors extracted from mice 3 weeks post treatment initiation with Control (Ctl) Ru1, gemcitabine (50mg/kg; twice per week) GEM or a combination of both (R+G). Values were normalized to ß-actin levels. * p < 0.05, **** p < 0.0001, ns = not significant, as determined by one-way ANOVA with Dunnett post-test, compared to untreated (Ctl). C-D Mean fold-change ± SEM in the expression of murine mtDNA-encoded genes mt-Atp6 and mt-Cox1 or the nuclear DNA encoded gene Drp1 in the heart (C) or liver (D) from mice harboring Panc265 orthotopic tumors, 3 weeks post treatment initiation with Control (C), Ru1 (R), gemcitabine (50mg/kg; twice per week) (G), or a combination of both (R+G). Values were normalized to Hprt levels. * p < 0.05, ** p < 0.01, **** p < 0.0001, ns = not significant, as determined by one-way ANOVA with Dunnett post-test, compared to untreated (Ctl). E WB analysis of mitochondria OXPHOS complex proteins using the Mitoprofile Total OXPHOS antibody cocktail in addition to GAPDH (loading control). Shown are bands corresponding to Complex (C)V, CIII, CII, CIV, and CI from Panc185 cells that were either untreated (CTL) or treated with Ru1 (100µM) or Ru1-met (100µM) for 48 h and 48 h + 24 h after removing treatment. F Mean fold-change ± SEM of mitochondrial complex bands determined in (E) by densitometric analysis and normalized to GAPDH. (n=4 pooled WBs, * p < 0.05, ** p < 0.01, ns = not significant, as determined by one-way ANOVA with Dunnett post-test, compared to untreated (CTL). G Mean fold-change ± SD in the mitochondria mass probe NAO in untreated (-), Ru1 (100µM) or Ru1-met (100µM)-treated Panc185 and PancA6L cells (48 h). * p < 0.05, **** p < 0.0001, ns = not significant, as determined by one-way ANOVA with Dunnett post-test, compared to untreated (Ctl)